Applied Biosystems Launches New Genomic Analysis Tools that Simplify Disease Research
16 Sept 2009Applied Biosystems has announced the launch of genomic analysis tools that will help researchers in studies that associate genetic variation with specific diseases. These new tools, known as the MeltDoctor™ High-Resolution Melt (HRM) Reagents, are for research use only and are part of a PCR-based workflow that enables researchers to cost-effectively scan DNA from a variety of biological sample types to detect the smallest genetic variations among samples.
The MeltDoctor HRM Reagents facilitate high resolution melt, or HRM analysis, which is a powerful technique for the detection of mutations, polymorphisms and epigenetic differences in double-stranded DNA samples. These reagents enable the characterization of variant nucleic acid sequences for advanced genomics applications such as mutation scanning, genotyping, and methylation analysis. These tools, including software, are part of an HRM analysis workflow optimized for Applied Biosystems real-time PCR systems that is simple to perform, requiring minimal quantities of both sample and reagent to capture a significantly greater amount of detail and information for variant types, without the complexity and expense of other methods.
HRM analysis is emerging as a technique of choice among genomics researchers and is one of several different analysis methods offered by the Genomics Core Lab at the Institute of Research in Immunology and Cancer at the University of Montreal. The IRIC Genomics Core Facility has been using an early-access version of the MeltDoctor HRM Reagents to simplify SNP analyses and to characterize new variants in candidate genes for immunology and cancer research studies.
Pierre Chagnon, Ph.D., Director of Genomics Core Facilities, IRIC, University of Montreal
“We have developed a melt analysis workflow for SNP analyses that includes the MeltDoctor HRM analysis reagents. This workflow helps us to deliver cost-effective, rapid genotyping projects for our core lab customers. The reagents are easy to use, as they do not require chemical optimization, and quickly generate accurate results for our customers the first time they submit their research samples.”
John Gerace, Vice President and General Manager of Life Technologies’ PCR Systems Business
“Applied Biosystems is dedicated to expanding the applications of our industry-leading portfolio of PCR tools for advanced genomics. The MeltDoctor High Resolution Melt Reagents significantly enhance HRM analysis, while broadening our offerings for technologies that contribute to the understanding of genetic variation for translational research applications that study the relationship between genomics and diseases.”
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MeltDoctor™ HRM Master Mix
Thermo Fisher ScientificHigh Resolution Melting (HRM) analysis is a post-PCR analysis method used to identify genetic variation in nucleic acid sequences. The MeltDoctor™ HRM Master Mix contains all components (excluding template and primers) formulated for superior HRM performance across a wide range of genomic targets. Unlike some competitive mixes, the MeltDoctor™ HRM Master Mix does not require additional mixing prior to use, and was developed and optimized solely for HRM applications. - Low background fluorescence - High brightness in the presence of double-stranded DNA - Minimal temperature shift of DNA melting due to dye binding - Thermal stability to tolerate PCR cycling conditions - No inhibition of polymerase activity, resulting in high PCR efficiency - A dNTP blend including dUTP, which minimizes carryover contamination by allowing amplicon degradation by uracil DNA glycosylase (UDG) in subsequent PCR reactions Perform Mutation Scanning HRM analysis can be used to scan for mutations in target genes for the identification of variant samples prior to sequencing analysis (Figure 1). As a mutation scanning technique, HRM offers significant advantages over conventional methods such as Denaturing High- Performance Liquid Chromatography (DHPLC) and Denaturing Gradient Gel Electrophoresis (DGGE). Specifically, the advantages of HRM for mutation scanning include: - Low reagent consumption, with little waste: HRM requires only a 20 µL PCR reaction for analysis of each sample, eliminating the need for HPLC solvents or DGGE gels - Simple, fast workflow: no additional instrumentation is required after PCR amplification. A high-resolution melt step can be simply added to the end of the PCR profile for immediate analysis - Fast optimization: unlike DHPLC, thermal optimization is not required - Low sample consumption: following HRM analysis, the PCR product can be used directly in a Sanger sequencing reaction Quality Components - Magnesium salts and other buffer components, precisely formulated to obtain optimal HRM results. - AmpliTaq Gold® 360 DNA Polymerase, a highly purified DNA polymerase that provides hot-start performance, minimizing non-specific product formation and enabling reactions to be set up at room temperature - MeltDoctor™ HRM Dye, a stabilized form of the fluorescent SYTO® 9 double-stranded nucleic acid stain developed by Molecular Probes.